Phenotypic changes of vascular smooth muscle cells in three dimensional collagen gel for the development of vascular graft was composed of biomaterials

Abstract

We studied phenotypic changes of vascular smooth muscle cells (SMC) in basic experiments “biomimetic grafts.” To evaluate suitable conditions of phenotypic changes, we cultured smooth muscle cells in three dimensional collagen gel cultures to which extracellular matrix was added, and then examined phenotypic changes by immunobiological methods. Monoclonal antibodies against myosin heavy chains isoform, SMemb., SMZ were used as biological marker of phenotype (contractile, synthetic type). Extracellular matrix, laminin, fibronectin and elastin and cell suspension (1.6×10⁵ cells/0.6ml) were added to 5% type I collagen solution and gel formation was made, and then SMC was cultured for 4-7 days in 5% CO₂ incubator (37°C). We extracted myosin with The Guba Straub solution and performed electrophoresis with SDS-PAGE, and Western blotting. Phenotype of SMC in three dimensional collagen gel was SMemb. positive (=synthetic type) with no phenotypic changes. However, the SMemb. positivity of SMC was inhibited by extracellular matrix, laminin, fibronectin (10μg) and elastin. This is important for studies of phenotypic changes of vascular SMC in three dimensional collagen gel.